antibody

Chk2, phosphorylated (Ser516) (Checkpoint Kinase 2, Rad 53)

C4201-32A

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

Chk2, phosphorylated (Ser516) (Checkpoint Kinase 2, Rad 53)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (1-3). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68) followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4,5). Indeed, after DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (5-7). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 on residues Thr383 and Thr387 in the activation loop of the kinase domain (8). Chk2 autophosphorylation at Ser516 is important for optimal Chk2 function, and a Ser516Ala mutant Chk2 is defective in IR-induced apoptosis (9). Applications: Suitable for use in Western Blot. Other applications not tested. Recommended Dilution: Western Blot: 1:1000 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot. Store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Hu

Synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser516 of human Chk2.

Detects endogenous levels of Chk2 only when phosphorylated at serine 516. The antibody does not cross-react with Chk2 phosphorylated at other sites. Species Crossreactivity: Human

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol.

100ul (10 Western mini-blots)

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: 100ul (10 Western mini-blots) Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser516 of human Chk2. Specificity: Detects endogenous levels of Chk2 only when phosphorylated at serine 516. The antibody does not cross-react with Chk2 phosphorylated at other sites. Species Crossreactivity: Human Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

WB

-20°C

(1) Allen, J.B. et al. (1994) Genes Dev. 8, 2401–2415. (2) Weinert, T.A. et al. (1994) Genes Dev. 8, 652–665. (3) Murakami, H. and Okayama, H. (1995) Nature 374, 817–819. (4) Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179–186. (5) Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389–10394. (6) Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762–765. (7) Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934–5936. (8) Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537–30541. (9) Wu, X. and Chen, J. (2003) J. Biol. Chem. 278, 36163–36168.

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